담관결찰 쥐에서 안지오텐신 길항제의 간섬유화 억제 효과
Inhibitory Effect of Angiotensin Blockade on Hepatic Fibrosis in Common Bile Duct-ligated Rats
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배경/목적: 안지오텐신은 일반적으로 섬유화 과정에 주요 역할을 하는 물질로 밝혀져 있는데, 간섬유화 과정에 핵심 역할을 하는 간성상세포(hepatic stellate cell)에도 안지오텐신 수용체가 존재하여, 안지오텐신이 간섬유화에 영향을 미치는 것으로 추정하고 있다. 이에 간경변 모델에서 안지오텐신 길항제가 간섬유화 억제에 효과적인지 알아보고자 하였다. 대상 및 방법: 300 g의 Sprague Dawley 쥐를 대상으로 총담관결찰(bile duct ligation, 이하 BDL)을 한 후 약물을 투여하지 않은 대조군과 captopril (angiotensin converting enzyme inhibitor) 또는 losartan (angiotensin II receptor antagonist)을 투여한 실험군으로 다음과 같이 나누었다(각 군 n=6). 1군으로 배정된 쥐들은 BDL 이후 아무 약제도 투여하지 않았으며, 2군에서는 BDL 이후 2주째부터 매일 100 mg/kg의 captopril을 물에 녹여 2주간 투여하였고, 3군에서는 BDL 직후부터 매일 100 mg/kg의 captopril을 4주간 계속 투여하였으며, 4군에서는 BDL 직후부터 losartan을 매일 10 mg/kg씩 4주간 계속 투여하였다. 대상 수술 동물은 4주 후에 희생시킨 후 Masson-trichrome 염색을 하여 간섬유화의 정도를 분석하였고(Batts & Ludwig score), alpha-smooth muscle actin에 대한 면역조직화학염색 후 화상 분석을 하여 간성상세포의 발현 정도를 비교 분석하였으며, hydroxyproline 농도를 통한 간섬유화의 정량 검사, real time PCR을 이용한 collagen, procollagen, TGF-beta의 발현 정도를 각 군에서 분석하였다. 결과: 간섬유화의 Batts & Ludwig 점수는 1군: 3.8, 2군: 3.0, 3군: 2.6, 4군: 2.6로 BDL군에 비해 BDL 직후부터 약물을 투여한 3군, 4군에서 낮았고, alpha-smooth muscle actin의 발현 정도(%)도 1군: 11.9, 2군: 10.9, 3군: 2.6, 4군: 1.1로 3군, 4군에서 낮았다(p<0.05). 간 hydroxyproline 농도(μg/g liver tissue)는 1군: 45.8, 2군: 33.6, 3군: 25.5, 4군: 18.0로 1군에 비해 3군, 4군에서 감소되어 있었고, collagen, procollagen, TGF-beta 발현 정도 역시 1군에 비해 3군, 4군에서 낮았다(p<0.05). 결론: 간경변증 유발 쥐모델에서 안지오텐신 전환효소 억제제와 안지오텐신 II 수용체 억제제와 같은 안지오텐신 길항제는 간섬유화 억제에 효과적이다.
Abstract
Backgrounds and Aims: Angiotensin receptors are found on hepatic stellate cells, which participate in hepatic fibrosis. Therefore, it is presumed that angiotensin has a role in hepatic fibrosis. The aim of this study was to evaluate the effects of angiotensin blockade on inhibition of hepatic fibrosis in cirrhotic rat model. Material and methods: Cirrhosis with portal hypertension was produced by common bile duct ligation (BDL) in the adult Sprague-Dawley rats. They were classified into 4 groups (each group n=6) as follows; G1: BDL without drug, G2: BDL+captopril 100 mg/kg/day beginning 2 weeks after BDL, G3: BDL+captopril 100 mg/kg/day, starting just after BDL, G4: BDL+losartan 10 mg/kg/day, starting just after BDL. After 4 weeks following BDL, hepatic fibrosis was histomorphologically analyzed by Batts & Ludwig score. Alpha smooth muscle actin by immunohistochemical stain, hydroxyproline contents of liver tissue by spectrophotometry and expression of collagen, procollagen, and TGF-beta by real-time PCR were measured. Results: Batts & Ludwig score were 3.8, 3.0, 2.6,and 2.6 in G1, G2, G3, and G4, respectively. The expression of alpha-SMA was significantly lower in G3 and G4 than in G1; 11.9%, 10.9%, 2.6%, and 1.1% in G1, G2, G3, and G4, respectively (p<0.05). The concentration of hydroxyproline (μg/g liver tissue) was lower in G3 and G4 compared with G1 (p<0.05). Also, the administration of angiotensin blockade just after BDL significantly reduced the expression of collagen, procollagen, and TGF-beta mRNA. Conclusions: Angiotensin blockades are effective in the prevention of hepatic fibrosis in BDL rats.
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